Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CDK4 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CDK4
Met1-Glu303
Accession # P11802

Specificity

Detects human and mouse CDK4 in direct ELISA.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse CDK4 Antibody

Detection of Human and Mouse CDK4 antibody by Western Blot.

Detection of Human and Mouse CDK4 by Western Blot.

Western blot shows lysates of A431 human epithelial carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, and NIH-3T3 mouse embryonic fibroblast cell line. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for CDK4 at approximately 34 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
CDK4 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

CDK4 in HeLa Human Cell Line.

CDK4 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

CDK4 in RAW264.7 cells.

CDK4 was detected in immersion fixed RAW264.7 cells using Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Human/Mouse CDK4 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cells, and MCF‑7 human breast cancer cells, loaded at 0.2 mg/mL. A specific band was detected for CDK4 at approximately 38 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human/Mouse CDK4 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line

Simple Western

20 µg/mL
Sample: HeLa human cervical epithelial carcinoma cells, and MCF‑7 human breast cancer cells

Western Blot

1 µg/mL
Sample: A431 human epithelial carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, and NIH‑3T3 mouse embryonic fibroblast cell line

Reviewed Applications

Read 1 review rated 4 using AF5254 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CDK4

CDK4 (cyclin-dependent kinase 4) is a 34 kDa member of the CMGC Ser/Thr protein kinase family of enzymes. Within the cell, pRb/retinoblastoma protein is normally unphosphorylated and suppresses cell growth at the G1 stage of the cell cycle. When mitosis is required, CDK4 is phosphorylated, forms a complex with cyclin D1, 2 or 3, and phosphorylates pRb/retinoblastoma protein. This inactivates pRb and cells initiate DNA synthesis. Human CDK4 is 303 amino acids (aa) in length. It contains one protein kinase domain (aa 6‑295) and an activating phosphorylation site at Thr172. There are three potential variants; one that shows an alternate start site at Met75, a second that shows a two aa substitution for aa 89‑211, and a third that shows a 23 aa substitution for aa 89‑303. Full-length human and mouse CDK4 are 95% aa identical.

Long Name

Cyclin-dependent Kinase 4

Alternate Names

CMM3, PSK-J3

Entrez Gene IDs

1019 (Human); 12567 (Mouse); 94201 (Rat)

Gene Symbol

CDK4

UniProt

P11802

Additional CDK4 Products

Product Documents for Human/Mouse CDK4 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human/Mouse CDK4 Antibody

For research use only

Related Research Areas

Cell Cycle
Intracellular Kinases

Citations for Human/Mouse CDK4 Antibody

Customer Reviews for Human/Mouse CDK4 Antibody (1)

4 out of 5
1 Customer Rating
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  • Human/Mouse CDK4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: OVCAR-3 human ovarian carcinoma cell line
    Species: Human
    Verified Customer | Posted 08/16/2017

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Protocols

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